obi-ontology
NTR: Clone-seq
editor preferred term: Clone-seq textual definition: A massively-parallel site-directed mutagenesis approach leveraging next-generation sequencing to rapidly and cost-effectively generate a large number of mutant alleles. One colony of each mutagenesis attempt is put into one pool and combine multiple pools through multiplexing for one Illumina sequencing run. definition source for the textual definition: http://journals.plos.org/plosgenetics/article?id=10.1371/journal.pgen.1004819 logical definition: sequencing assay --> Clone-seq
I'm wondering if this is more of an experimental design, rather than a truly novel subtype of sequencing assay. The goal of "Clone-Seq" is "to rapidly and cost-effectively generate a large number of mutant alleles", rather than simply to assay a particular single sample for some feature of that sample. The equipment used to perform the sequencing aspects of "Clone-seq" is standard Illumina equipment. The novel portions are combining site-directed mutagenesis techniques and protein-protein interactome information to generate mutants, then organizing those mutants across standard high-throughput sequencing runs.
I wasn't asked, but I agree :) Also, the definition could be a bit more specific: Is this specific for bacterial cloning ('colony' makes me think that)? Is there something specific about the colony picking? That might make it a specific kind of assay...
On Mon, Oct 15, 2018 at 9:39 AM Dan Berrios [email protected] wrote:
@jahilton https://github.com/jahilton Do you agree?
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Yes, @DanBerrios, I agree with your characterization of the method. And yes, @bpeters42, site-directed mutagenesis, in general, uses a host cell (most often E. coli, I believe) as the vector. The distinction with Clone-seq, specifically, is the pooling/multiplexing aspect